Cent agreement for NREM epochs. There was only a main effect of data file (F(4, 16) = 28.53, p < 0.001). C,PLOS ONE | DOI:10.1371/journal.pone.0152473 March 31,34 /Endocannabinoid Signaling Regulates Sleep StabilityResults for percent agreement for REM epochs. There was only a main effect of data file (F(4, 16) = 33.18, p < 0.001). D, Shows data collapsed across scorers and the results of a paired comparison by data file. Grey, connected points superimposed on the bar graph indicate mean human:human and computer:human agreement for each data file. A paired t-test was performed for each vigilance state. Percent agreement was not significantly different for human: human vs human:computer scored data for wake (t(4) = 1.94, p = 0.12), NREM (t(4) = 1.09, p 0.34), or REM (t(4) = 0.77, p = 0.48). (PDF) S4 Fig. MAGL Inhibition with JZL184 Administration Before the LP Attenuates Gamma Frequency Oscillations During Sleep. A-C, Average power spectra for epochs of different vigilance states across the entire LP (left hand) and DP (right hand). Solid lines denote means and shaded region around lines denotes SEM. A, Wake. B, NREM. C, REM. D-F, Change over the day in summated power in different frequency bandwidths from the power spectra: delta (left hand column), theta (middle column), and gamma (right hand column). D, Wake epochs. Left panel: For wake delta, there was an overall interaction (treatment x time of day within photoperiod, F(24, 261.30) = 2.08, p = 0.003), Chaetocin web nested interaction (time of day within photoperiod, F(6, 242.10) = 9.33, p < 0.001), and a main effect of photoperiod (F(1, 134.75) = 6.88, p = 0.010). The only time point that significantly deviated from vehicle was during the first 3 Hr of the recovery day, when there was an increase in delta power (t(215.67) = 2.86, p fpsyg.2016.01448 31.18, p < 0.001), and main effects of both treatment (F (4, 163.42) = 22.79, p < 0.001) and photoperiod (F(1, 126.56) = 230.95, p < 0.001). Overall, JZL had dose-dependent effects on NREM gamma, with decreases observed following 8.0 (t (145.58) = -3.45, p = 0.003) and 16.0 mg/kg doses (t(94.93) = -4.97, p < 0.001). 8.0 mg/kg JZL decreased NREM gamma power for 18 Hr following drug administration (ZT 00?8: t(179.23) -2.71, p 0.030), and 16.0 mg/kg JZL reduced NREM gamma across the entire recording (ZT 00?0: t(116.56) -2.97, p 0.015).Cent agreement for NREM epochs. There was only a main effect of data file (F(4, 16) = 28.53, p < 0.001). C,PLOS ONE | DOI:10.1371/journal.pone.0152473 March 31,34 /Endocannabinoid Signaling Regulates Sleep StabilityResults for percent agreement for REM epochs. There was only a main effect of data file (F(4, 16) = 33.18, p < 0.001). D, Shows data collapsed across scorers and the results of a paired comparison by data file. Grey, connected points superimposed on the bar graph indicate mean human:human and computer:human agreement for each data file. A paired t-test was performed for each vigilance state. Percent agreement was not significantly different for human: human vs human:computer scored data for wake (t(4) = 1.94, p = 0.12), NREM (t(4) = 1.09, p 0.34), or REM (t(4) = 0.77, p = 0.48). (PDF) S4 Fig. MAGL Inhibition with JZL184 Administration Before the LP Attenuates Gamma Frequency Oscillations During Sleep. A-C, Average power spectra for epochs of different vigilance states across the entire LP (left hand) and DP (right hand). Solid lines denote means and shaded region around lines denotes SEM. A, Wake. B, NREM. C, REM. D-F, Change over the day in summated power in different frequency bandwidths from the power spectra: delta (left hand column), theta (middle column), and gamma (right hand column). D, Wake epochs. Left panel: For wake delta, there was an overall interaction (treatment x time of day within photoperiod, F(24, 261.30) = 2.08, p = 0.003), nested interaction (time of day within photoperiod, F(6, 242.10) = 9.33, p < 0.001), and a main effect of photoperiod (F(1, 134.75) = 6.88, p = 0.010). The only time point that significantly deviated from vehicle was during the first 3 Hr of the recovery day, when there was an increase in delta power (t(215.67) = 2.86, p SART.S23506 = 0.018). Middle panel: No effect of JZL184 on wake theta power. Right panel: For wake gamma power, there was a nested interaction (time of day within photoperiod, F(6, 253.19) = 6.08, p < 0.001) and main effects of both treatment (F(4,67.43) = 3.21, p = 0.018) and photoperiod (F(1, 179.51) = 115.90, p < 0.001). Specifically, 16 mg/kg JZL reduced gamma power during the first 3 Hr of the LP (ZT 00?3: t(57.18) = -2.68, p = 0.038). E, NREM epochs. Left panel: For NREM delta power, there was no effect of JZL treatment. Middle panel: For NREM theta power, there was an overall interaction (treatment x time of day within photoperiod, F(24, 268.23) = 1.64, p = 0.033), a nested interaction (time of day within photoperiod, F(6, 238.31) = 20.36, p < 0.001), and a main effect of photoperiod (F(1, 159.84) = 85.90, p < 0.001). However, there were no specific time points where JZL184 significantly altered NREM theta power relative to vehicle. Right panel: For NREM gamma power, there was an overall interaction (treatment x time of day within photoperiod, F(24, 267.36) = 2.46, p < 0.001), a nested interaction (time of day within photoperiod, F(6, 234.97) = fpsyg.2016.01448 31.18, p < 0.001), and main effects of both treatment (F (4, 163.42) = 22.79, p < 0.001) and photoperiod (F(1, 126.56) = 230.95, p < 0.001). Overall, JZL had dose-dependent effects on NREM gamma, with decreases observed following 8.0 (t (145.58) = -3.45, p = 0.003) and 16.0 mg/kg doses (t(94.93) = -4.97, p < 0.001). 8.0 mg/kg JZL decreased NREM gamma power for 18 Hr following drug administration (ZT 00?8: t(179.23) -2.71, p 0.030), and 16.0 mg/kg JZL reduced NREM gamma across the entire recording (ZT 00?0: t(116.56) -2.97, p 0.015).